title: [Nuclear localization of oligonucleotides decoy effect on nuclear factor-kappaB activity].
abstract: To investigate the effect of the localization of oligonucleotides decoy (ODNs decoy) on the activation of nuclear factor-kappaB (NF-kappaB) in TNF-alpha induced HeLa cells. The mercapto group-modified nuclear localization signal (NLS) peptide was covalently conjugated to amino group-modified NF-kappaB ODNs decoy by Sulfo-SMCC cross-linker. The NLS-ODNs decoy was transfected into HeLa cells by TransME transfection reagent. The intracellular distribution of fluorescent labeled NLS-ODNs decoy was detected with a microscope. The cell viability was detected by MTT assay, and then the activity of NF-kappaB in cell nuclear extract was assayed by electrophoretic mobility shift assay (EMSA). The results showed that NLS peptide was successfully conjugated to ODNs decoy by Sulfo-SMCC cross-linker. The NLS-ODNs decoy effectively entered into nucleus with high rate of 17.9%. It was observed that the cell viability of HeLa cell was not significantly affected by the transfection of NLS-ODNs decoy, while NLS-ODNs decoy significantly inhibited the activation of NF-kappaB in TNF-alpha induced HeLa cells nuclear extracts. This experiment can provide a new covalent conjugation of NLS peptide to ODNs can effectively drive decoy into nucleus, and thus improve its inhibitory effects on the activation a transcription factor.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Yingxun
authors.initials: Y
authors.lastname: Liu
authors.forename: Fusheng
authors.initials: F
authors.lastname: Quan
authors.forename: Jinke
authors.initials: J
authors.lastname: Wang
authors.forename: Xueyao
authors.initials: X
authors.lastname: Bai
mesh_terms: D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism*; D019913:Nuclear Localization Signals / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D016328:NF-kappa B; D019913:Nuclear Localization Signals; D009841:Oligonucleotides